Direct immunoblotting from histological sections of brain onto nitrocellulose: evaluation with the protein neurophysin.
A method for the isolation and localization of proteins and peptides from histological sections of rat and human brain by immunoblotting is described. For validation, the well-characterized protein neurophysin was electrophoretically transferred from formaldehyde-fixed or fresh tissue sections onto a nitrocellulose membrane. Neurophysin on the nitrocellulose membrane was detected by a specific antibody reaction. The antibody against neurophysin was visualized either by using secondary antibodies, conjugated with peroxidase or by protein A gold, followed by enhancement with silver. With this simple and fast method, neurophysin (or other proteins and peptides) can be identified on nitrocellulose membranes in areas that correspond to anatomically defined regions. Since the procedure combines the advantages of precise regional localization of polypeptides with the specificity of antibody-antigen reactions, the method may prove useful for rapid screening of the distribution of peptides or proteins in (brain) tissue.
Reisinger
P W
PW
Department of Anatomy, University of Munich, Federal Republic of Germany.
Unger
J W
JW
D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry; D006801:Humans; D007031:Hypothalamus / Q000737:chemistry* / Q000166:cytology; D008297:Male; D008851:Micropore Filters; D009481:Neurophysins / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains
D009481:Neurophysins